Abstract
BackgroundEpithelio- and endotheliotropic viruses often exert polarized entry and release that may be responsible for viral spread and dissemination. Hantaviruses, mostly rodent-borne members of the Bunyaviridae family infect epithelial and endothelial cells of different organs leading to organ dysfunction or even failure. Endothelial and renal epithelial cells belong to the target cells of Old World hantavirus. Therefore, we examined the release of hantaviruses in several renal epithelial cell culture models. We used Vero cells that are commonly used in hantavirus studies and primary human renal epithelial cells (HREpC). In addition, we analyzed MDCKII cells, an epithelial cell line of a dog kidney, which represents a widely accepted in vitro model of polarized monolayers for their permissiveness for hantavirus infection.ResultsVero C1008 and primary HREpCs were grown on porous-support filter inserts for polarization. Monolayers were infected with hantavirus Hantaan (HTNV) and Puumala (PUUV) virus. Supernatants from the apical and basolateral chamber of infected cells were analyzed for the presence of infectious particles by re-infection of Vero cells. Viral antigen and infectious particles of HTNV and PUUV were exclusively detected in supernatants collected from the apical chamber of infected Vero C1008 cells and HREpCs. MDCKII cells were permissive for hantavirus infection and polarized MDCKII cells released infectious hantaviral particles from the apical surface corresponding to the results of Vero and primary human epithelial cells.ConclusionsPathogenic Old World hantaviruses are released from the apical surface of different polarized renal epithelial cells. We characterized MDCKII cells as a suitable polarized cell culture model for hantavirus infection studies.
Highlights
Epithelio- and endotheliotropic viruses often exert polarized entry and release that may be responsible for viral spread and dissemination
Release of hantaviruses HTNV and PUUV from the apical surface of Vero C1008 cells and primary human renal epithelial cells (HREpC) To examine the release of hantaviruses, Vero C1008 cells were cultivated on membrane inserts and infected with HTNV and PUUV by apical inoculation
These results demonstrate that pathogenic Old World hantavirus HTNV and PUUV are released from the apical surface of Vero C1008 cells
Summary
Epithelio- and endotheliotropic viruses often exert polarized entry and release that may be responsible for viral spread and dissemination. Endothelial and renal epithelial cells belong to the target cells of Old World hantavirus. We analyzed MDCKII cells, an epithelial cell line of a dog kidney, which represents a widely accepted in vitro model of polarized monolayers for their permissiveness for hantavirus infection. For the Old World hantaviruses HTNV and PUUV we could show that the viruses enter Vero and HUVE cells via the apical surface [17]. We examined the release of the pathogenic Old World hantaviruses PUUV and HTNV from polarized renal epithelial cells. Since MDCKII cells represent a well established and widely used model for polarized epithelial monolayers [18,19], we investigated the susceptibility of MDCKII cells as a possible cell culture model for hantavirus infection and analyzed entry and release of hantaviruses in this cell line
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